【作者】 袁子民；

【导师】 贾天柱；

【作者基本信息】 辽宁中医药大学 ， 生药学， 2006， 博士

【摘要】 本课题立足于国家“八五”攻关课题“常用中药饮片肉豆蔻研究”（85920-08），结合国家“十五”重大科技专项“创新药物和中药现代化”——肉豆蔻炮制工艺及质量标准规范化研究课题（2001BA701A55-29）的技术要求，对肉豆蔻炮制方法中存在的争议问题、炮制目的、炮制的传统和现代文献、化学成分、药理毒理、炮制工艺、质量标准等方面，进行了深入系统的研究，比较了炮制前后的化学成分的质与量的变化、药理作用的差异性等，从而揭示肉豆蔻的炮制原理，对肉豆蔻炮制工艺的规范化、临床合理用药及质量标准建立等，均具有重要的指导意义。 通过古代和现代炮制文献研究，归纳总结了肉豆蔻的多种炮制方法及其炮制目的。在此基础上对多种炮制方法进行比较研究，为确立科学合理的炮制方法提供了研究思路。 在肉豆蔻化学成分研究中，从肉豆蔻生品中分离并鉴定了9个化合物，分别为去氢二异丁香酚（dehydrodiisoeugenol）、利卡啉-B（licarin-B）、β-谷甾醇、胡萝卜苷、黄樟醚（safrol）、肉豆蔻醚（myristicin）、榄香脂素（elemicin）、甲基丁香酚、甲基异丁香酚。并通过化学分离、药理跟踪试验，确定了肉豆蔻中的止泻成分为甲基丁香酚、甲基异丁香酚。 在肉豆蔻炮制工艺研究中，采用L₉（3⁴）正交试验，以肉豆蔻醚、黄樟醚、甲基丁香酚、甲基异丁香酚含量变化为指标，进行综合评分，优选出肉豆蔻最佳炮制工艺为水浸泡6hr，在170～180℃麸煨25min，每100kg肉豆蔻用麦麸40kg。并进行中试放大实验验证，结果工艺合理、可行。同时对肉豆蔻炮制前后的挥发油成分进行GC-MS分析，结果肉豆蔻经炮制后挥发油中成分发生了质和量的变化，其中炮制后有13个新成分增加，4个成分消失，且止泻成分甲基丁香酚、甲基异丁香酚含量增加，毒性成分肉豆蔻醚、黄樟醚含量降低。 本研究完成了肉豆蔻质量标准及麸煨肉豆蔻质量标准及起草说明。建立了肉豆蔻中肉豆蔻醚、黄樟醚、甲基丁香酚、甲基异丁香酚、去氢二异丁香酚的含量测定方法。并进行了显微鉴别、薄层鉴别、水分、灰分、农药残留、重金属及有害元素等测定。 采用HPLC法对肉豆蔻炮制前后甲醇提取物、挥发油图谱比较，结果甲醇提取物的化学成分的谱图变化并不太明显，但某些主要化学成分的含量有明显变化；炮制后挥发油中甲基丁香酚、甲基异丁香酚含量增加，同时又有两个未知成分增加，丁香酚、异丁香酚肉豆蔻醚、黄樟醚含量降低。同时采用计算机辅助解析技术对肉豆蔻炮制前后的IR图谱进行了对比研究，结果各炮制品的相关系数差异性很大，均低于0.983，说明炮制对其整体化学成分均有一定影响。变化较大的为润后麸煨和单蒸炮制品。 对商品肉豆蔻饮片及其炮制品HPLC指纹图谱的构建进行了初步探讨，所建立的肉豆蔻饮片及炮制品HPLC指纹图谱中，以去氢二异丁香酚为参照物峰，标定了7个共有特征（指纹）峰，各特征峰的调整保留时间之比和相对峰面积可以作为肉豆蔻质量优劣的依据，同时为肉豆蔻进口药材的质量标准及其炮制品研究提供资料。 提取生品肉豆蔻挥发油，对挥发油进行不同温度加热一定时间，采用HPLC法测定不同温度加热后挥发油中甲基丁香酚、甲基异丁香酚含量，与未加热的原油进行含量动态变化分析。结果生品肉豆蔻挥发油开始加热后随着温度的升高逐渐增加，当温度超过200±10℃时，开始明显增加，至240±10℃时达到最大值，然后随温度升高逐渐降低。肉豆蔻炮制后止泻成分甲基丁香酚、甲基异丁香酚的增加，可能主要是由挥发油受热后某些成分发生更多还原

【Abstract】 The subject of the processing principle of Nutmeg was a section of the National tackle key problem in eighth five years plan named as "study of Traditional Chinese herbal pieces of Nutmeg "and the National important technical subject item in tenth five years plan named as "innovation and Modernization of Traditional Chinese Medicine". The author of this dissertation studied the processing technology, chemical substances, pharmacology and toxicology activities, the quality standard of Nutmeg systematically. This research explored the processing principle of Nutmeg.Nine chemicals were isolated, purified and identified from Nutmeg. They were deihydrodiisoengenol, licarin-B, daucosterol, safrol, myristicin, elemicin, methyleugenol, methylisoeugenol. Methyleugenol and methylisoeugenol had active action of anti-diarrhea.The optimal processing method was studied by orthogonal design with the contents of myristicin, safrol, methyleugenol, methylisoeugenol as indicators. The results showed that the optimal processing condition was: Nutmeg was soaked with water for 6hr, then roasted with wheat bran at 170-180°C for 25min. GC-MS method was developed to determine the constituents of volatile oil from Nutmeg and the processed products. The experiments showed that the constituents of volatile oil from Nutmeg after processing had qualitative and quantitative difference. 13 new compounds occurred, but 4 compounds disappeared; the contents of methyleugenol ?and methylisoeugenol, the anti-diarrhea constituents, increased; and the contents of myristicin and safrol, toxic constituents, decreased.A HPLC methanol was developed to determine the contents of myristicin. safrol, methyleugenol, methylisoeugenol, dihydrodiisoengenol.A HPLC method was developed to compare the chromatograph of the methanoi extraction and volatile oil from Nutmeg and the processing products. The results indicated that the chromatograph of the menthol extraction had no significant difference, but the contents of some main chemical constituents had remarkable difference. The contents of methyleugenol, methylisoeugenol and two uncertain compounds in volatile oil from the processing products increased. But the contents of engenol, isoengenol, myristicin, safrol decreased. Computer Aided Analysis was developed to analysis the difference of IR chromatograph between Nutmeg and the processed products. The results showed that the correlative coefficient of different processing methods of Nutmeg, all below 0.983, had obvious difference. The results showed that the processing methods had some effects on the whole chemical constituents.The chemical fingerprints of Nutmeg and the processing products were更多还原